HK1074976B - Yeast extract that enhances soup stock flavor - Google Patents

Description

Yeast extract for enhancing flavor of broth

Technical Field

The present invention refers to a yeast extract which can optimally enhance all of the flavors of broth (soupstock) while enhancing the body and mixed flavor of broth prepared by boiling animal meat or marine foods, thereby sufficiently improving the deliciousness thereof.

Background

In various cookings, so-called "soups" are used, which are prepared by decocting animal meats or marine foods. Thus, the meat has special flavor, and the flavor is stronger and has mixed flavor. However, when it is intended to apply the rich flavor of the broth to the meal, concentrating the broth becomes important. Disadvantageously, this makes the broth expensive. Therefore, the broth is generally used in a state of flavor enhancer by adding so-called flavor enhancer such as sodium glutamate, sodium guanylate, sodium inosinate, or mixed Hydrolyzed Vegetable Protein (HVP), Hydrolyzed Animal Protein (HAP), yeast extract, or the like.

Prior to the filing of this application, examples of known patent documents include: japanese patent publication (Kokai) No. 6-113789A (1994), "Yeast extract containing a large amount of palatable nucleotides and production thereof; "Japanese patent publication (Kokai) No. 1-112966A (1989)," preparation of novel yeast extract ", and Japanese patent publication (Kokoku) No. 4-58945B (1992)," preparation of yeast extract for improving taste quality ".

However, the previously mentioned techniques only impart a certain taste to the broth. Although all flavor titers could be improved, there was no emphasis on the thick and mixed flavors inherent in broths. As a result, the taste of the broth disadvantageously becomes unbalanced.

Summary of The Invention

It is an object of the present invention to provide a yeast extract which is capable of preferably enhancing all the flavors of a stock while enhancing the inherent body and mixed flavor of the stock, thereby sufficiently enhancing the deliciousness thereof.

The present inventors have conducted intensive studies in order to achieve this object. As a result of the studies, it was found that when the peptide component having a molecular weight of 10,000 or more is contained in at least 10% in the yeast extract, the body and the mixed flavor of the broth can be particularly increased. The present inventors have further found that when the peptide component having a molecular weight of 2,000 or less is present in the yeast extract in an amount of at least 55%, a yeast extract can be obtained which is capable of preferably increasing the body, the mixed flavor of the broth without deteriorating all the taste qualities of the broth.

In the yeast extract production process, when monosodium glutamate or succinic acid is produced and mixed into the yeast extract, the resultant is used together with a general broth, which preferably only increases the inherent taste of the broth. While balancing astringent, pungent and unpleasant tastes without the emphasis on prominent taste components, these findings have led to the completion of the present invention.

More specifically, the present invention relates to:

(1) a yeast extract comprising at least 10% of the total peptides detected, said peptides having a molecular weight of 10,000 or greater, wherein said peptides are detected by the following method: obtaining yeast extract by digesting or degrading yeast, filtering through a membrane with a pore size of 1um, filtering the filtrate with a gel, and detecting the absorbance of the eluate fraction at 220 nm;

(2) the yeast extract as described in (1), wherein the peptides having a molecular weight of 2000 or less account for at least 55% of the total amount of the peptides to be detected;

(3) the yeast extract as described in (1) or (2), which contains at least 10% by solid of monosodium glutamate; and

(4) the yeast extract as in (1) or (2), which contains at least 0.6% succinic acid on a solid basis.

The present invention will be described in detail below.

The yeast extract of the present invention is prepared by digesting or degrading yeast. The yeast extract is filtered through a membrane having a pore size of 1um, and the peptide fraction having a molecular weight of 10,000 or more accounts for at least 10% of the total amount of the filtered peptides. The yeast extract is not particularly limited by the production method, the type of yeast or the source thereof. For example, the yeast extract can be obtained by collecting yeast cells by a commonly used culture method, washing the yeast cells to make a suspension in a sufficient amount of water, removing the yeast extract by autolysis or using yeast cell wall digesting enzymes, adding exoproteases instead of endoproteases to degrade proteins, and preparing the yeast extract into a liquid, paste or powder by a conventional technique. Yeast cells that accumulate large amounts of glutamate or succinate may also be used.

The yeast extract obtained in this way, having a unique molecular weight distribution, for example, a component consisting of amino acids, proteins, or peptides, can be separated by membrane filtration with a pore size of 1um (hereinafter referred to as "peptide mode").

Generally, this peptide pattern structure is analyzed by gel filtration, and is continuously fractionated according to molecular weight. Peptides can be fractionated by this technique and their absorbance at 220nm detected, thereby determining the peptide pattern.

Yeast extract was used as a carrier for gel filtration and analyzed by the following method using peptide fractionation using Superdex peptide HR10/20 column (Pharmacia Biotech Co.). Specifically, the yeast extract was dissolved in 0.02M disodium hydrogenphosphate-sodium dihydrogenphosphate buffer containing 0.25M NaCl in a buffer liquid nitrogen content of about 0.05%, and the solution was filtered through a filter having a pore size of 1 μ M to obtain a sample. The sample was gel filtered through a Superdex peptide HR10/20 column (Pharmacia Biotech) for peptide fractionation, and the absorbance at 220nm was measured. Thus, the ratio of the components of the yeast extract was determined by molecular weight fractionation.

The yeast extract of the present invention is added to a stock, particularly to a stock mainly prepared from marine foods or animal meats, and can enhance the flavor of all the stocks most appropriately, and also sufficiently improve the deliciousness, while giving the stock a rich flavor and a mixed taste. The yeast extract can enhance the thick flavor and mixed taste of the stock because its composition is very similar to the average peptide pattern of chicken broth, beef broth and dried bonito broth (hereinafter referred to as "average peptide pattern of stock"). The soup-stock has an average peptide pattern characterized by a peptide fraction having a molecular weight of 10,000 or more being at least 10% of the total amount of peptides and a peptide fraction having a molecular weight of 2000 or less being at least 55% of the total amount of peptides.

More specifically, the peptide pattern of the yeast extract of the invention resembles the average peptide pattern of a soup-stock. In this way, the yeast extract is compatible with these soups, and its taste is also well mixed with the soups. At the same time, the yeast extract of the present invention has a strong flavor and a taste of a mixture significantly better than those of the existing yeast extracts or other seasonings. Accordingly, when the yeast extract is combined with the broth, the yeast extract can most suitably enhance the flavor of the broth.

Further, monosodium glutamate (hereinafter abbreviated as "MSG") or succinic acid is sometimes produced at the same time as the production of the yeast extract. When MSG is present in solid form in an amount of 10% or more of the yeast extract product or, likewise, succinic acid is present in solid form in an amount of 0.6% or more of the yeast extract product, it may be further preferred to enhance the flavor of the yeast extract alone while balancing the astringent, pungent and unpleasant tastes without emphasizing prominent taste components. This affects the body taste and flavor mix, particularly in the peptide mode, provided by components having a molecular weight of 10,000 or greater, as well as the effect. Even though the yeast extract product contains a large amount of flavor-enhancing substances, these flavors are not prominent.

Components that do not pass through a filter with a pore size of 1um are generally only considered as doubts. Depending on their function, yeast extracts contaminated with these components are undesirable for taste and other reasons.

Preferred embodiments of the invention

The present invention will be described in detail below with reference to specific examples, but the scope of the present invention is not limited to only the following examples.

Example 1

A suspension containing 10% of solids was prepared by culturing Saccharomyces cerevisiae 3-2-C-7, FERM BP-10022, deposited in the national Institute of Advanced Industrial science and Technology, in a 300-liter fermenter by a conventional technique by adding water to yeast cells to prepare a suspension containing 10% of solids, adjusting the pH of the cell suspension to 5.7 with a 40% NaOH solution, adding cell wall digestive enzyme YL-15(Amano enzyme preparation Co.) at a solid content of 0.1%, reacting at 45 ℃ for 8 hours and 10 hours, respectively, after completion of the reaction, the reaction product was heated at 70 ℃ for 30 minutes to inactivate the enzyme, and then cooled to 40 ℃, after which peptidase R (Amano enzyme preparation Co.) was added at a solid content of 0.2%, reacted at 40 ℃ for 2 hours, and after completion of the reaction, the reaction product was heated at 70 ℃ for 30 minutes, inactivating enzyme to obtain pasty yeast extract by conventional method.

The obtained yeast extract was dissolved in 0.02M disodium hydrogenphosphate-sodium dihydrogenphosphate buffer solution containing 0.25M NaCl in a buffer liquid nitrogen content of about 0.05%, and the solution was filtered through a filter membrane having a pore size of 1um to obtain a sample. The sample was gel filtered through a Superdex peptide HR10/20 column (Pharmacia Biotech) for peptide fractionation, and the absorbance at 220nm was measured. Thus, the ratio of the yeast extract components was determined by molecular weight fractionation based on the obtained area ratio.

Table 1 shows the molecular weight fractions of the yeast extracts obtained. In the extract, the reaction time was 8 hours and 10 hours, and the ratio of peptides having a molecular weight of 10,000 or more was 14.9% and 10.2%, respectively. The ratio of amino acids and peptides having a molecular weight of 2,000 or less was 73.9% and 79.0%, respectively.

Table 1 shows the results of analysis of yeast extract components of the present invention in comparison with the ratios determined by molecular weight fractionation and the commercial yeast extracts.

Table 1: ingredient ratio of commercial yeast extract classified according to molecular weight

	≤2,000	2,000 to 5,000	5,000 to 10,000	≥10,000
					Yeast extract of the present invention (reaction time 8 hours)	73.9	8.4	2.9	14.9
Yeast extract of the present invention (reaction time 10 hours)	79.0	8.6	2.5	10.2
					Aromild	90.4	2.8	1.3	5.5
Yeast extract HR	88.2	5.3	2.1	4.4
					Yeast extract H	84.5	11.6	1.5	1.5
Ajipulse BF	78.3	11.7	2.0	8.0
					Super yeast extract	81.0	7.3	2.8	8.9
Meast S	84.9	10.6	2.0	2.5
					Meast N	88.7	8.5	1.3	1.5
Yeast extract 21-TF	79.7	15.1	2.5	2.7

Aromild, Ajipule BF (KOHJIN Co., Ltd.), Yeast extract HR (Takeda-Kirin food Co., Ltd.); yeast extract H (Kyowa Hakko Kogyo co., ltd.); super yeast extract (Ajinomoto, ltd.); meast S, Meast N (Asahi Food & Healthcare limited); yeast extract 21-TF (Nicotiana japonica Co.).

Each yeast extract was added to a dried bonito soup at an amount of 0.5%, and the function thereof was examined to evaluate the enhancement effect on the flavor of the stock soup. The results are shown in table 2.

Table 2: enhancing soup stock flavor

	The soup storage agent has effect in enhancing flavor
		Is free of	0
Yeast extract of the present invention (reaction time 8 hours)	6
		Yeast extract of the present invention (reaction time 10 hours)	5
Aromild	2
		Yeast extract HR	2
Yeast extract H	1
		Ajipulse BF	3
Super yeast extract	3
		Meast S	2
Meast N	1
		Yeast extract 21-TF	2

^*High values show higher enhancement.

As the number of components having a molecular weight of 10,000 or more increases, the effect of enhancing the flavor of the broth is also higher. The yeast extract of the present invention (reaction time: 8 hours), which contains 14.9% of this component, is very effective. It is apparent from Table 1 that, in order to achieve optimum effects, components having a molecular weight of 10,000 or more should account for at least 10% of the total amount.

Example 2

Saccharomyces bayanus cells (a variant of Saccharomyces cerevisiae HG-1, 3-2-C-7 cells, producing large amounts of glutamic acid) were cultured by conventional techniques using a 300-liter fermentor. Water was added to the yeast cells to prepare a suspension containing 10% solids, and the pH of the cell suspension was adjusted to 5.7 with 40% NaOH solution. Cell wall digesting enzyme YL-15(Amano enzyme preparation Co.) was added at a solid content of 0.1% and reacted at 45 ℃ for 8 hours. After the reaction was completed, the reaction product was heated at 70 ℃ for 30 minutes to inactivate the enzyme, and then cooled to 40 ℃. Thereafter, peptidase R (Amano enzyme preparation Co.) was added at a solid content of 0.2% and reacted at 40 ℃ for 2 hours. After the reaction was completed, the reaction product was heated at 70 ℃ for 30 minutes to inactivate the enzyme, and then the product was heated at 70 ℃ for another 30 minutes to obtain a paste-like yeast extract by the conventional technique.

AG-1 cells producing a large amount of glutamic acid were obtained as follows.

10ml of the solution is added⁵-10⁶Saccharomyces baeckii cell (Saccharomyces cerevisiae 3-2-C-7) The suspension was placed in a petri dish and irradiated with uv light for 60 seconds (uv lamp at 15 watts, distance: 30 cm) while stirring with a stirrer. The yeast suspension irradiated with ultraviolet rays was diluted sufficiently, and the diluted suspension was inoculated on YPD agarose medium plates, cultured at 30 ℃ for 2 days to grow colonies, and the colonies were transferred on YPP agarose medium plates using pyruvic acid as a carbon source. After further culturing at 30 ℃ for 2 days, cells in which colonies did not grow were isolated. The separated cells which do not take up pyruvic acid were inoculated into 5ml YPD liquid medium with a platinum ring, cultured at 30 ℃ under shaking at 300rpm for 24 hours, and centrifuged at 3000rpm for 10 minutes to obtain cells. The glutamic acid content of the cells was thus determined.

YPD agarose culture medium

Yeast extract 0.5%

2.0 percent of peptone

Glucose 4.0%

Potassium dihydrogen phosphate 0.5%

Magnesium sulfate 2.0%

Agarose 2.0%

YPD liquid culture medium

Yeast extract 0.5%

2.0 percent of peptone

Glucose 4.0%

Potassium dihydrogen phosphate 0.5%

Magnesium sulfate 2.0%

YPP agarose medium

Yeast extract 0.5%

2.0 percent of peptone

Pyruvic acid 1.0%

Potassium dihydrogen phosphate 0.5%

Magnesium sulfate 2.0%

Agarose 2.0%

The glutamic acid content was measured by the following method.

2ml of water was added to the cells, and the resultant was reacted at 70 ℃ for 30 minutes, centrifuged again, and 1ml of the supernatant was collected after centrifugation and filtered through a 0.45um filter to obtain a sample solution for measurement. The resultant was analyzed by Hitachi L-8500 amino acid analyzer. Based on the quantification of glutamate, the total amount of MSG was determined.

HG-1 cells containing more MSG than the parent cell (3-2-C-7) were sorted out.

The sorted cells were compared with the yeast extract of example 1, and the results are shown in Table 3. The sorted cells were added to 5% heated meat extract at a relative level of 0.5% to compare the flavor enhancing effect, and the results are shown in Table 4.

Table 3: yeast extract component ratio determined based on molecular weight fractionation and MSG content in solid component

	≤ 2,000	2,000 to 5,000	5,000 to 10,000	≥10,000	MSG
						Example 1 Yeast extract	73.9	8.4	2.9	14.9	7.5
Example 2 Yeast extract	75.6	8.0	2.5	15.2	12.5

Table 4: enhancing soup stock flavor

	Enhancing soup stock flavor
		Example 1 Yeast extract	6
Example 2 Yeast extract	7

The results show that the effect of enhancing the flavor of the broth is further enhanced when a large amount of MSG is produced in the yeast extract.

A sodium glutamate reagent (Wako Pure chemical industries, Ltd.) was added to the yeast extract obtained in example 1 to evaluate the effect of enhancing the flavor of broth.

Table 5 shows the evaluation of MSG content and the effect of enhancing the flavour of the broths.

TABLE 5

MSG content	7.5％	8％	9％	10％	11％
						Enhancing soup stock flavor	6	6	6	7	7

As is apparent from table 5, when the MSG content is 10% or more, the effect of enhancing the flavor of the broth is enhanced.

Example 3

A300-liter fermentor was used to grow Saccharomyces bayanus cells (Saccharomyces cerevisiae 3-2-C-7) by conventional techniques, water was added to the yeast cells to prepare a suspension containing 10% solids, and the pH of the cell suspension was adjusted to 5.7 with 40% NaOH solution. Cell wall digesting enzyme YL-15(Amano enzyme preparation Co.) was added at a solid content of 0.1% and reacted at 45 ℃ for 8 hours. After the reaction was completed, the reaction product was heated at 70 ℃ for 30 minutes to inactivate the enzyme, and then cooled to 45 ℃. Thereafter, Neutrase F3G (Amano enzyme preparations) having a weaker self-exonuclease activity was added at a solid content of 0.2% and reacted at 45 ℃ for 1 hour and 2 hours, respectively. After the reaction was completed, the reaction product was heated at 70 ℃ for 30 minutes to inactivate the enzyme, and a paste yeast extract was obtained by the conventional technique.

The component ratios of the obtained yeast extract, determined by molecular weight fractionation, were compared with those of the yeast extract obtained in example 1, and the results are shown in Table 6. The effect of increasing the flavor of the dried bonito soup was compared, and the results are shown in Table 7.

Table 6: determining the ratio of the components of yeast extract according to molecular weight classification

	≤ 2,000	2,000 to 5,000	5,000 to 10,000	≥ 10,000
					Example 1 Yeast extract	73.9	8.4	2.9	14.9
Enzyme reaction: 2 hours	56.8	15.6	12.4	15.2
					Enzyme reaction: 1 hour	45.2	21.2	17.9	15.7

Table 7: enhancing soup stock flavor

	Flavor enhancing effect
		Example 1 Yeast extract	6
Enzyme reaction: 2 hours	6
		Enzyme reaction: 1 hour	5

The yeast extract having a molecular weight of 2,000 or less in an amount of 45.2% has less effect of increasing the flavor of the broth after 1 hour of the enzymatic reaction than the yeast extract having an amount of 55% in an amount of such components.

Example 4

Saccharomyces bayanus cells (a variant of Saccharomyces cerevisiae KY-2, 3-2-C-7 cells, producing large amounts of succinic acid) were cultured by conventional techniques using a 300-liter fermentor. Water was added to the yeast cells to prepare a suspension containing 10% solids, and the pH of the cell suspension was adjusted to 5.7 with 40% NaOH solution. Cell wall digesting enzyme YL-15(Amano enzyme preparation Co.) was added at a solid content of 0.1% and reacted at 45 ℃ for 8 hours. After the reaction was completed, the reaction product was heated at 70 ℃ for 30 minutes to inactivate the enzyme, and then cooled to 40 ℃. Thereafter, peptidase R (Amano enzyme preparation Co.) was added at a solid content of 0.1% and reacted at 40 ℃ for 1 hour. After the reaction was completed, the reaction product was heated at 70 ℃ for 30 minutes to inactivate the enzyme, and a paste yeast extract was obtained by the conventional technique.

KY-2 cells producing large amounts of succinic acid were obtained as follows.

10ml of the solution is added⁵-10⁶A suspension of cells of Saccharomyces bayanus (Saccharomyces cerevisiae 3-2-C-7) was placed in a petri dish, irradiated with ultraviolet rays for 60 seconds (ultraviolet lamp, distance: 50 cm), and stirred with a stirrer. Fully diluting the yeast suspension irradiated by ultraviolet rays, inoculating the diluted suspension onto a YPD agarose medium plate, culturing at 30 ℃ for 2 days to grow colonies, transferring the colonies onto an YPP medium plate using pyruvic acid as a carbon source, continuously culturing at 30 ℃ for 2 days, and separating cells without growing the colonies. The separated cells which did not take up pyruvic acid were inoculated into 5ml of YPD liquid medium with a platinum ring, cultured at 30 ℃ under shaking at 300rpm for 1 day, and centrifuged at 3000rpm for 10 minutes to obtain cells. The succinic acid content of the cells was thus determined.

The succinic acid content was measured by the following method.

2ml of water was added to the cells, and the resultant was reacted at 70 ℃ for 30 minutes, centrifuged again, and 1ml of the supernatant was collected after centrifugation and filtered through a 0.45um filter. The sample solution was analyzed by high performance liquid chromatography (DGU-14A, Shimadzu).

Model DGU-14A

Pillar SHODEX RSpak KC-811

Guard column SHODEX RSpak KC-G

Column temperature 40 deg.C

Eluent 3mM perchloric acid 1.0ml/min

ST3R 0.5 diluted with 1/10 reaction solution 0.5ml/min

Detection of UV 430nm

The component ratios of the obtained yeast extract determined by molecular weight fractionation were compared with those of the yeast extract obtained in example 1, and the results are shown in Table 8. The results of comparison of the effect of increasing the flavor of dried bonito soup are shown in Table 9.

Table 8: the component ratio of the obtained yeast extract determined by molecular weight fractionation and succinic acid in the solid component

	≤ 2,000	2,000 to 5,000	5,000 to 10,000	≥10,000	Succinic acid
						Example 1 Yeast extract	73.9	8.4	2.9	14.9	0.32
Example 4 Yeast extract	74.3	7.9	2.7	15.1	1.26

Table 9: enhancing soup stock flavor

	Enhancing soup stock flavor
		Example 1 Yeast extract	6
Example 4 Yeast extract	7

The results show that the effect of enhancing the flavor of the broth is further enhanced when a large amount of succinic acid is produced in the yeast extract.

Succinic acid (Wako Pure chemical industries, Ltd.) was added to the yeast extract of example 1 to evaluate the effect of enhancing the flavor of the broth.

Table 10 shows the evaluation of succinic acid content and the effect of enhancing the flavor of the broth.

Watch 10

Succinic acid content	0.32％	0.5％	0.6％	0.7％	10.0％
						Enhancing soup stock flavor	6	6	7	7	7

As is apparent from table 10, when the succinic acid content is 0.6% or more, the effect of enhancing the flavor of the broth is enhanced.

Industrial applicability

The present invention provides a yeast extract which can enhance optimally all the flavors of soup-stock while enhancing the inherent body and mixed flavor of soup-stock, thereby sufficiently improving the deliciousness thereof.

Claims (4)

1. A yeast extract in which peptides having a molecular weight of 10,000 or more account for at least 10% of the total of all peptides tested, said peptides being tested by the following method: yeast extract is obtained by digesting or degrading yeast, filtered with a filter membrane having a pore size of 1um, the filtrate is subjected to gel filtration, and the eluate fraction is analyzed for absorbance at 220 nm.

2. The yeast extract according to claim 1, wherein the peptides having a molecular weight of 2,000 or less account for at least 55% of the total amount of all peptides tested.

3. The yeast extract of claim 1 or 2, comprising at least 10% monosodium glutamate on a solids basis.

4. The yeast extract of claim 1 or 2, comprising at least 0.6% succinic acid on a solids basis.